4-Substituted Tetracyclines and Methods of Use Thereof

ABSTRACT

The present invention pertains, at least in part, to novel substituted tetracycline compounds. These tetracycline compounds can be used to treat numerous tetracycline compound-responsive states, such as bacterial infections and neoplasms, as well as other known applications for tetracycline compounds such as blocking tetracycline efflux and modulation of gene expression.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.11/258,613, filed Oct. 25, 2005, which claims the benefit of U.S.Provisional Application No. 60/622,027, filed on Oct. 25, 2004 and U.S.Provisional Application No. 60/673,827, filed on Apr. 21, 2005, thecontents of each of which is incorporated herein by reference.

BACKGROUND OF THE INVENTION

The development of the tetracycline antibiotics was the direct result ofa systematic screening of soil specimens collected from many parts ofthe world for evidence of microorganisms capable of producingbacteriocidal and/or bacteriostatic compositions. The first of thesenovel compounds was introduced in 1948 under the name chlortetracycline.Two years later, oxytetracycline became available. The elucidation ofthe chemical structure of these compounds confirmed their similarity andfurnished the analytical basis for the production of a third member ofthis group in 1952, tetracycline. A new family of tetracyclinecompounds, without the ring-attached methyl group present in earliertetracyclines, was prepared in 1957 and became publicly available in1967; and minocycline was in use by 1972.

Recently, research efforts have focused on developing new tetracyclineantibiotic compositions effective under varying therapeutic conditionsand routes of administration. New tetracycline analogues have also beeninvestigated which may prove to be equal to or more effective than theoriginally introduced tetracycline compounds. Examples include U.S. Pat.Nos. 2,980,584; 2,990,331; 3,062,717; 3,165,531; 3,454,697; 3,557,280;3,674,859; 3,957,980; 4,018,889; 4,024,272; and 4,126,680. These patentsare representative of the range of pharmaceutically active tetracyclineand tetracycline analogue compositions.

Historically, soon after their initial development and introduction, thetetracyclines were found to be highly effective pharmacologicallyagainst rickettsiae; a number of gram-positive and gram-negativebacteria; and the agents responsible for lymphogranuloma venereum,inclusion conjunctivitis, and psittacosis. Hence, tetracyclines becameknown as “broad spectrum” antibiotics. With the subsequent establishmentof their in vitro antimicrobial activity, effectiveness in experimentalinfections, and pharmacological properties, the tetracyclines as a classrapidly became widely used for therapeutic purposes. However, thiswidespread use of tetracyclines for both major and minor illnesses anddiseases led directly to the emergence of resistance to theseantibiotics even among highly susceptible bacterial species bothcommensal and pathogenic (e.g., pneumococci and Salmonella). The rise oftetracycline-resistant organisms has resulted in a general decline inuse of tetracyclines and tetracycline analogue compositions asantibiotics of choice.

SUMMARY OF THE INVENTION

In one embodiment, the invention includes tetracycline compounds offormula (I):

wherein:

R^(2′) and R^(2″) are each independently hydrogen, alkyl, alkenyl,alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino,arylalkyl, aryl, heterocyclic, heteroaromatic or a prodrug moiety;

R¹⁰, R¹¹ and R¹² are each independently hydrogen, alkyl, aryl, benzyl,arylalkyl, or a pro-drug moiety;

R^(3′) is hydroxyl, hydrogen, or a pro-drug moiety;

R⁴ is O or N—OR^(4a);

R^(4a) is hydrogen, alkyl, alkenyl, alkynyl, or aryl;

R⁵ and R^(5′) are each independently hydroxyl, hydrogen, thiol,alkanoyl, aroyl, alkaroyl, aryl, heteroaromatic, alkyl, alkenyl,alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino,arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy;

R⁶ and R^(6′) are each independently hydrogen, methylene, absent,hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy,alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;

R⁷ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl,alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,arylalkyl, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(7c))₀₋₁C(═W′)WR^(7a);

R⁸ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl,alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,alkylamino, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(8c))₀₋₁C(=E′)ER^(8a);

R⁹ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl,alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,arylalkyl, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(9c))₀₋₁CC(═Z′)ZR^(9a);

R^(7a), R^(7b), R^(7c), R^(7d), R^(7e), R^(7f), R^(8a), R^(8b), R^(9c),R^(8d), R^(8e), R^(8f), R^(9a), R^(9b), R^(9c), R^(9d), R^(9e), andR^(9f) are each independently hydrogen, acyl, alkyl, alkenyl, alkynyl,alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl,aryl, heterocyclic, heteroaromatic or a prodrug moiety;

R¹³ is hydrogen, hydroxy, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,aryl, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;

E is CR^(8d)R^(8e), S, NR^(8b) or O;

E′ is O, NR^(8f), or S;

W is CR^(7d)R^(7e), S, NR^(7b) or O;

W′ is O, NR^(7f), or S;

X is CHC(R¹³Y′Y), C═CR¹³Y, CR^(6′)R⁶, S, NR⁶, or O;

Y′ and Y are each independently hydrogen, halogen, hydroxyl, cyano,sulfhydryl, amino, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;

Z is CR^(9d)R^(9e), S, NR^(9b) or O;

Z′ is O, S, or NR^(9f), and pharmaceutically acceptable salts, estersand enantiomers thereof.

In another embodiment, the invention pertains, at least in part, totetracycline compounds of formula (II):

wherein:

R^(2′) and R^(2″) are each independently hydrogen, alkyl, alkenyl,alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino,arylalkyl, aryl, heterocyclic, heteroaromatic or a prodrug moiety;

R¹⁰, R¹¹ and R¹² are each independently hydrogen, alkyl, aryl, benzyl,arylalkyl, or a pro-drug moiety;

R^(3′) is hydroxyl, hydrogen, or a pro-drug moiety;

R⁴ is NR^(4c)R^(4d);

R^(4c) and R^(4d) are each independently hydrogen, sulfonyl,arylcarbonyl, arylaminocarbonyl, aryloxycarbonyl, alkylcarbonyl,alkylaminocarbonyl, alkyloxycarbonyl, acyl, alkyl, alkenyl, alkynyl, oraryl;

R⁵ and R^(5′) are each independently hydroxyl, hydrogen, thiol,alkanoyl, aroyl, alkaroyl, aryl, heteroaromatic, alkyl, alkenyl,alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino,arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy;

R⁶ and R^(6′) are each independently hydrogen, methylene, absent,hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy,alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;

R⁷ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl,alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,arylalkyl, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(7c))₀₋₁C(═W′)WR^(7a);

R⁸ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl,alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,alkylamino, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(8c))₀₋₁C(=E′)ER^(8a);

R⁹ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl,alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,arylalkyl, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(9c))₀₋₁C(═Z′)ZR^(9′);

R^(7a), R^(7b), R^(7c), R^(7d), R^(7e), R^(7f), R^(8a), R^(8b), R^(8c),R^(8d), R^(8e), R^(8f), R^(9a), R^(9b), R^(9c), R^(9d), R^(9e), andR^(9f) are each independently hydrogen, acyl, alkyl, alkenyl, alkynyl,alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl,aryl, heterocyclic, heteroaromatic or a prodrug moiety;

R¹³ is hydrogen, hydroxy, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,aryl, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;

E is CR^(8d)R^(8e), S, NR^(8b) or O;

E′ is O, NR^(8f), or S;

W is CR^(7d)R^(7e), S, NR^(7b) e or O;

W′ is O, NR^(7f), or S;

X is CHC(R¹³Y′Y), C═CR¹³Y, CR^(6′)R⁶, S, NR⁶, or O;

Y′ and Y are each independently hydrogen, halogen, hydroxyl, cyano,sulfhydryl, amino, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;

Z is CR^(9d)R^(9e), S, NR^(9b) or O;

Z′ is O, S, or NR^(9f), and pharmaceutically acceptable salts, estersand enantiomers thereof.

In another further embodiment, the invention pertains, at least in part,to methods for treating subjects for tetracycline responsive states byadministering to them an effective amount of a tetracycline compound ofthe invention, e.g., a compound of Formula I or II or a tetracyclinecompound otherwise described herein.

In another further embodiment, the invention pertains, at least in part,to to pharmaceutical compositions which comprise an effective amount ofa tetracycline compound of the invention, e.g., a compound of Formula Ior II or a tetracycline compound otherwise described herein, and apharmaceutically acceptable carrier.

DETAILED DESCRIPTION OF THE INVENTION

The present invention pertains, at least in part, to novel substitutedtetracycline compounds. These tetracycline compounds can be used totreat numerous tetracycline compound-responsive states, such asbacterial infections and neoplasms, as well as other known applicationsfor minocycline and tetracycline compounds in general, such as blockingtetracycline efflux and modulation of gene expression.

The term “tetracycline compound” includes many compounds with a similarring structure to tetracycline. Examples of tetracycline compoundsinclude: chlortetracycline, oxytetracycline, demeclocycline,methacycline, sancycline, chelocardin, rolitetracycline, lymecycline,apicycline; clomocycline, guamecycline, meglucycline, mepylcycline,penimepicycline, pipacycline, etamocycline, penimocycline, etc. Otherderivatives and analogues comprising a similar four ring structure arealso included (See Rogalski, “Chemical Modifications of Tetracyclines,”the entire contents of which are hereby incorporated herein byreference). Table 1 depicts tetracycline and several known othertetracycline derivatives.

TABLE 1

Oxytetracycline

Demeclocycline

Minocycline

Methacycline

Doxycycline

Chlortetracycline

Tetracycline

Sancycline

Chelocardin

Other tetracycline compounds which may be modified using the methods ofthe invention include, but are not limited to,6-demethyl-6-deoxy-4-dedimethylaminotetracycline; tetracyclino-pyrazole;7-chloro-4-dedimethylaminotetracycline;4-hydroxy-4-dedimethylaminotetracycline;12α-deoxy-4-dedimethylaminotetracycline;5-hydroxy-6α-deoxy-4-dedimethylaminotetracycline;4-dedimethylamino-12α-deoxyanhydrotetracycline;7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline;tetracyclinonitrile; 4-oxo-4-dedimethylaminotetracycline 4,6-hemiketal;4-oxo-11a C1-4-dedimethylaminotetracycline-4,6-hemiketal;5a,6-anhydro-4-hydrazon-4-dedimethylamino tetracycline;4-hydroxyimino-4-dedimethylamino tetracyclines;4-hydroxyimino-4-dedimethylamino 5a,6-anhydrotetracyclines;4-amino-4-dedimethylamino-5a,6 anhydrotetracycline;4-methylamino-4-dedimethylamino tetracycline;4-hydrazono-11a-chloro-6-deoxy-6-demethyl-6-methylene-4-dedimethylaminotetracycline; tetracycline quaternary ammonium compounds;anhydrotetracycline betaines; 4-hydroxy-6-methyl pretetramides; 4-ketotetracyclines; 5-keto tetracyclines; 5a, 11a dehydro tetracyclines; 11aC1-6, 12 hemiketal tetracyclines; 11a C1-6-methylene tetracyclines; 6,13diol tetracyclines; 6-benzylthiomethylene tetracyclines; 7,11a-dichloro-6-fluoro-methyl-6-deoxy tetracyclines; 6-fluoro(α)-6-demethyl-6-deoxy tetracyclines; 6-fluoro (β)-6-demethyl-6-deoxytetracyclines;6-αacetoxy-6-demethyl tetracyclines; 6-βacetoxy-6-demethyl tetracyclines; 7,13-epithiotetracyclines;oxytetracyclines; pyrazolotetracyclines; 11a halogens of tetracyclines;12a formyl and other esters of tetracyclines; 5,12a esters oftetracyclines; 10,12a-diesters of tetracyclines; isotetracycline;12-a-deoxyanhydro tetracyclines;6-demethyl-12a-deoxy-7-chloroanhydrotetracyclines; B-nortetracyclines;7-methoxy-6-demethyl-6-deoxytetracyclines;6-demethyl-6-deoxy-5a-epitetracyclines; 8-hydroxy-6-demethyl-6-deoxytetracyclines; monardene; chromocycline; 5a methyl-6-demethyl-6-deoxytetracyclines; 6-oxa tetracyclines, and 6 thia tetracyclines.

4-Substituted Tetracycline Compounds

In one embodiment, the invention includes tetracycline compounds offormula (I)

wherein:

R^(2′) and R^(2″) are each independently hydrogen, alkyl, alkenyl,alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino,arylalkyl, aryl, heterocyclic, heteroaromatic or a prodrug moiety;

R¹⁰, R¹¹ and R¹² are each independently hydrogen, alkyl, aryl, benzyl,arylalkyl, or a pro-drug moiety;

R^(3′) is hydroxyl, hydrogen, or a pro-drug moiety;

R⁴ is O or N—OR^(4a);

R^(4a) is hydrogen, alkyl, alkenyl, alkynyl, or aryl;

R⁵ and R^(5′) are each independently hydroxyl, hydrogen, thiol,alkanoyl, aroyl, alkaroyl, aryl, heteroaromatic, alkyl, alkenyl,alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino,arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy;

R⁶ and R^(6′) are each independently hydrogen, methylene, absent,hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy,alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;

R⁷ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl,alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,arylalkyl, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(7c))₀₋₁C(═W′)WR^(7a);

R⁸ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl,alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,alkylamino, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(8c))₀₋₁C(=E′)ER^(8a);

R⁹ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl,alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,arylalkyl, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(9c))₀₋₁C(═Z′)ZR^(9a);

R^(7a), R^(7b), R^(7c), R^(7d), R^(7e), R^(7f), R^(8a)R^(8b), R^(8c),R^(8d), R^(8e), R^(8f), R^(9a), R^(9b), R^(9c), R^(9d), R^(9e), andR^(9f) are each independently hydrogen, acyl, alkyl, alkenyl, alkynyl,alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl,aryl, heterocyclic, heteroaromatic or a prodrug moiety;

R¹³ is hydrogen, hydroxy, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,aryl, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;

E is CR^(8d)R^(8e), S, NR^(8b) or O;

E′ is O, NR^(8f), or S;

W is CR^(7d)R^(7e), S, NR^(7b) or O;

W′ is O, NR^(7f), or S;

X is CHC(R¹³Y′Y), C═CR¹³Y, CR^(6′)R⁶, S, NR⁶, or O;

Y′ and Y are each independently hydrogen, halogen, hydroxyl, cyano,sulfhydryl, amino, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;

Z is CR^(9d)R^(9e), S, NR^(9b) or O;

Z′ is O, S, or NR^(9f), and pharmaceutically acceptable salts, estersand enantiomers thereof.

In a further embodiment, R^(2′), R³, R¹⁰, R¹¹, and R¹² are each hydrogenor a prodrug moiety; X is CR⁶R^(6′); and R^(2″), R⁵, R^(5′), R⁶, andR^(6′) are each hydrogen.

Alternatively, R⁵ and R^(5′) are hydrogen and X is CR⁶R^(6′), wherein R⁶is methyl and R^(6′) is hydroxy. Alternatively, when R⁵ is hydroxyl; Xis CR⁶R^(6′); R⁶ is methyl; and R^(5′) and R^(6′) are hydrogen. In yetanother embodiment, X is CR⁶R^(6′); R⁵, R^(5′), R⁶ and R^(6′) arehydrogen atoms and R⁷ is dimethylamino.

In one embodiment, R⁹ is hydrogen. In another embodiment, R⁹ issubstituted or unsubstituted aryl (e.g., phenyl or heteroaryl). Inanother embodiment, R⁹ is substituted or unsubstituted alkyl. In afurther embodiment, R⁹ is aminoalkyl, e.g., aminomethyl (e.g.,—CH₂—N^(9n)R^(9m), wherein R^(9n) is hydrogen or a prodrug and R^(9m) ishydrogen or lower alkyl).

In another embodiment, R⁷ is hydrogen. In yet another embodiment, R⁷ issubstituted or unsubstituted aryl, e.g., phenyl or heteroaryl (e.g.,pyridinyl, pyrrolyl, pyrazinyl, etc.). In another embodiment, R⁷ issubstituted or unsubstituted amino (e.g., dimethylamino), nitro orhalogen.

In a further embodiment, R⁴ is O. In another embodiment, R⁴ is N—OH.

In another further embodiment, the tetracycline compound is:

In a further embodiment, the invention pertains to compounds of theformula (II):

wherein:

R^(2′) and R^(2″) are each independently hydrogen, alkyl, alkenyl,alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino,arylalkyl, aryl, heterocyclic, heteroaromatic or a prodrug moiety;

R¹⁰, R¹¹ and R¹² are each independently hydrogen, alkyl, aryl, benzyl,arylalkyl, or a pro-drug moiety;

R^(3′) is hydroxyl, hydrogen, or a pro-drug moiety;

R⁴ is NR^(4c)R^(4d);

R^(4c) and R^(4d) are each independently hydrogen, sulfonyl,arylcarbonyl, arylaminocarbonyl, aryloxycarbonyl, alkylcarbonyl,alkylaminocarbonyl, alkyloxycarbonyl, acyl, alkyl, alkenyl, alkynyl, oraryl;

R⁵ and R^(5′) are each independently hydroxyl, hydrogen, thiol,alkanoyl, aroyl, alkaroyl, aryl, heteroaromatic, alkyl, alkenyl,alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino,arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy;

R⁶ and R^(6′) are each independently hydrogen, methylene, absent,hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy,alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;

R⁷ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl,alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,arylalkyl, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(7c))₀₋₁C(═W′)WR^(7a);

R⁸ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl,alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,alkylamino, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(8c))₀₋₁C(=E′)ER^(8a);

R⁹ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl,alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,arylalkyl, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(9c))₀₋₁C(═Z′)ZR^(9a);

R^(7a), R^(7b), R^(7c), R^(7d), R^(7e), R^(7f), R^(8a), R^(8b), R^(8c),R^(8d), R^(8e), R^(8f), R^(9a), R^(9b), R^(9c), R^(9d), R^(9e), andR^(9f) are each independently hydrogen, acyl, alkyl, alkenyl, alkynyl,alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl,aryl, heterocyclic, heteroaromatic or a prodrug moiety;

R¹³ is hydrogen, hydroxy, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,aryl, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;

E is CR^(8d)R^(8e), S, NR^(8b) or O;

E′ is O, NR^(8f), or S;

W is CR^(7d)R^(7e), S, NR^(7b) or O;

W′ is O, NR^(7f), or S;

X is CHC(R¹³Y′Y), C═CR¹³Y, CR^(6′)R⁶, S, NR⁶, or O;

Y′ and Y are each independently hydrogen, halogen, hydroxyl, cyano,sulfhydryl, amino, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;

Z is CR^(9d)R^(9e), S, NR^(9b) or O;

Z′ is O, S, or NR^(9f), and pharmaceutically acceptable salts, estersand enantiomers thereof.

In a further embodiment, R^(2′), R³, R¹⁰, R¹¹, and R¹² are each hydrogenor a prodrug moiety; X is CR⁶R^(6′); and R^(2″), R⁵, R^(5′), R⁶, andR^(6′) are each hydrogen.

Alternatively, R⁵ and R^(5′) are hydrogen and X is CR⁶R^(6′), wherein R⁶is methyl and R^(6′) is hydroxy. Alternatively, when R⁵ is hydroxyl; Xis CR⁶R^(6′); R⁶ is methyl; and R^(5′) and R^(6′) are hydrogen. In yetanother embodiment, X is CR⁶R^(6′); R⁵, R^(5′), R⁶ and R^(6′) arehydrogen atoms and R⁷ is dimethylamino.

In one embodiment, R⁹ is hydrogen. In another embodiment, R⁹ issubstituted or unsubstituted aryl (e.g., phenyl or heteroaryl). Inanother embodiment, R⁹ is substituted or unsubstituted alkyl. In afurther embodiment, R⁹ is aminoalkyl, e.g., aminomethyl (e.g.,—CH₂—N^(9n)R^(9m), wherein R^(9n) is hydrogen or a prodrug and R^(9m) ishydrogen or lower alkyl).

In another embodiment, R⁷ is hydrogen. In yet another embodiment, R⁷ issubstituted or unsubstituted aryl, e.g., phenyl or heteroaryl (e.g.,pyridinyl, pyrrolyl, pyrazinyl, etc.). In another embodiment, R⁷ issubstituted or unsubstituted amino (e.g., dimethylamino), nitro orhalogen. In another embodiment, R⁸ is hydrogen.

In another embodiment, R^(4c) is hydrogen. In a further embodiment,R^(4d) is also hydrogen. Alternatively, R^(4d) is substituted orunsubstituted arylcarbonyl, arylaminocarbonyl, substituted orunsubstituted arylsulfonyl or alkylcarbonyl.

In another embodiment, the tetracycline compound is selected from thegroup consisting of:

In one embodiment, the tetracycline compounds of the invention do notinclude those described in U.S. Ser. No. 09/660,598, 09/823,884,09/852,908, 10/819,343, 10/820,456, 09/894,805, 09/895,796, 09/895,812,09/895,797, 09/895,857, 10/097,634, 10/759,484, 10/337,914, 10/636,437,10/752,378, or 10/740,961. The entire contents of each of theseapplications are hereby incorporated herein in their entirety.

In another embodiment, the tetracycline compounds of the invention donot include tetracycline compounds wherein R⁷ is selected from the groupconsisting of hydrogen, amino, nitro, mono(lower alkyl)amino, halogen,di(lower alkyl)amino, ethoxythiocarbonylthio, azido, acylamino,diazonium, cyano, and hydroxyl; R^(6′) is selected from the groupconsisting of hydrogen and methyl; R⁶ and R⁵ are selected from the groupconsisting of hydrogen and hydroxyl; R⁴ is selected from the groupconsisting of NOH, N—NH-A, and NH-A, where A is a lower alkyl group; R⁸is selected from the group consisting of hydrogen and halogen; R⁹ isselected from the group consisting of hydrogen, amino, azido, nitro,acylamino, hydroxy, ethoxythiocarbonylthio, mono(lower alkyl)amino,halogen, di(lower alkyl)amino and RCH(NH₂)CO; R is hydrogen or loweralkyl; and pharmaceutically acceptable and unacceptable salts thereof.In another embodiment, the invention does not include the compoundsdescribed in U.S. Pat. No. 6,506,740, incorporated herein by referencein its entirety.

Methods for Synthesizing Tetracycline Compounds of the Invention

The tetracycline compounds of this invention can be synthesized usingthe methods described in the Schemes, Examples, and/or by othertechniques known to those of ordinary skill in the art.

The substituted tetracycline compounds of the invention can besynthesized using the methods described in the following schemes and byusing art recognized techniques. All novel substituted tetracyclinecompounds described herein are included in the invention as compounds.

9- and 7-substituted tetracyclines can be synthesized by the methodshown in Scheme 1. As shown in Scheme 1, 9- and 7-substitutedtetracycline compounds can be synthesized by treating a tetracyclinecompound (e.g., doxycycline, 1A), with sulfuric acid and sodium nitrate.The resulting product is a mixture of the 7-nitro and 9-nitro isomers(1B and 1C, respectively). The 7-nitro (1B) and 9-nitro (1C) derivativesare treated by hydrogenation using hydrogen gas and a platinum catalystto yield amines 1D and 1E. The isomers are separated at this time byconventional methods. To synthesize 7- or 9-substituted alkenylderivatives, the 7- or 9-amino tetracycline compound (1E and 1F,respectively) is treated with HONO, to yield the diazonium salt (1G and1H). The salt (1G and 1H) is treated with an appropriate reactivereagent to yield the desired compound (e.g., in Scheme 1,7-cyclopent-1-enyl doxycycline (1H) and 9-cyclopent-1-enyl doxycycline(1I)).

As shown in Scheme 2, tetracycline compounds of the invention wherein R⁷is a carbamate or a urea derivative can be synthesized using thefollowing protocol. Sancycline (2A) is treated with NaNO₂ under acidicconditions forming 7-nitro sancycline (2B) in a mixture of positionalisomers. 7-nitrosancycline (2B) is then treated with H₂ gas and aplatinum catalyst to form the 7-amino sancycline derivative (2C). Toform the urea derivative (2E), isocyanate (2D) is reacted with the7-amino sancycline derivative (2C). To form the carbamate (2G), theappropriate acid chloride ester (2F) is reacted with 2C.

As shown in Scheme 3, tetracycline compounds of the invention, whereinR⁷ is a heterocyclic (i.e. thiazole) substituted amino group can besynthesized using the above protocol. 7-amino sancycline (3A) is reactedwith Fmoc-isothiocyanate (3B) to produce the protected thiourea (3C).The protected thiourea (3C) is then deprotected yielding the activesancycline thiourea (3D) compound. The sancycline thiourea (3D) isreacted with an α-haloketone (3E) to produce a thiazole substituted7-amino sancycline (3F).

7-alkenyl tetracycline compounds, such as 7-alkynyl sancycline (4A) and7-alkenyl sancycline (4B), can be hydrogenated to form 7-alkylsubstituted tetracycline compounds (e.g., 7-alkyl sancycline, 4C).Scheme 4 depicts the selective hydrogenation of the 7-position double ortriple bond, in saturated methanol and hydrochloric acid solution with apalladium/carbon catalyst under pressure, to yield the product.

In Scheme 5, a general synthetic scheme for synthesizing 7-position arylderivatives is shown. A Suzuki coupling of an aryl boronic acid with aniodosancycline compound is shown. An iodo sancycline compound (5B) canbe synthesized from sancycline by treating sancycline (5A) with at leastone equivalent N-iodosuccinimide (NIS) under acidic conditions. Thereaction is quenched, and the resulting 7-iodo sancycline (5B) can thenbe purified using standard techniques known in the art. To form the arylderivative, 7-iodo sancycline (5B) is treated with an aqueous base(e.g., Na₂CO₃) and an appropriate boronic acid (5C) and under an inertatmosphere. The reaction is catalyzed with a palladium catalyst (e.g.,Pd(OAc)₂). The product (5D) can be purified by methods known in the art(such as HPLC). Other 7-aryl, alkenyl, and alkynyl tetracyclinecompounds can be synthesized using similar protocols.

The 7-substituted tetracycline compounds of the invention can also besynthesized using Stille cross couplings. Stille cross couplings can beperformed using an appropriate tin reagent (e.g., R—SnBu₃) and ahalogenated tetracycline compound, (e.g., 7-iodosancycline). The tinreagent and the iodosancycline compound can be treated with a palladiumcatalyst (e.g., Pd(PPh₃)₂Cl₂ or Pd(AsPh₃)₂Cl₂) and, optionally, with anadditional copper salt, e.g., CuI. The resulting compound can then bepurified using techniques known in the art.

The compounds of the invention can also be synthesized using Heck-typecross coupling reactions. As shown in Scheme 6, Heck-typecross-couplings can be performed by suspending a halogenatedtetracycline compound (e.g., 7-iodosancycline, 6A) and an appropriatepalladium or other transition metal catalyst (e.g., Pd(OAc)₂ and CuI) inan appropriate solvent (e.g., degassed acetonitrile). The substrate, areactive alkene (6B) or alkyne (6D), and triethylamine are then addedand the mixture is heated for several hours, before being cooled to roomtemperature. The resulting 7-substituted alkenyl (6C) or 7-substitutedalkynyl (6E) tetracycline compound can then be purified using techniquesknown in the art.

To prepare 7-(2′-Chloro-alkenyl)-tetracycline compounds, the appropriate7-(alkynyl)-sancycline (7A) is dissolved in saturated methanol andhydrochloric acid and stirred. The solvent is then removed to yield theproduct (7B).

As depicted in Scheme 8, 5-esters of 9-substituted tetracyclinecompounds can be formed by dissolving the 9-substituted compounds (8A)in strong acid (e.g. HF, methanesulphonic acid, andtrifluoromethanesulfonic acid) and adding the appropriate carboxylicacid to yield the corresponding esters (8B).

As shown in Scheme 9 below, 7 and 9 aminomethyl tetracyclines may besynthesized using reagents such as hydroxymethyl-carbamic acid benzylester.

A method for derivatizing tetracycline compounds at the 4 position hasbeen discovered through chemical modification via oxidation of the C4position to produce a C4 oxime moiety. The oxime moiety is reduced to aC4 amino group as shown in Scheme 10:

In one embodiment, the invention pertains to a method for synthesizing4-substituted tetracycline compounds. The method includes contacting atetracycline compound with an effective amount of an oxidizing agent toform a 4-oximesubstituted tetracycline compound, and contacting the4-oximesubstituted tetracycline compound with a reducing agent, suchthat the 4-aminosubstituted tetracycline compound is formed.

The term “oxidizing agent” includes agents which are capable ofoxidizing the C4 dialkylamine group to an oxime moiety. In oneembodiment, the oxidizing agent is hydroxylamine.

Reducing agents are also known in the art. Examples of reducing agentsare described in Comprehensive Organic Transformations (“COT”) 2^(nd)Ed., Larock, 304, 305, incorporated herein by reference. In oneembodiment, the reducing agent is palladium on carbon with hydrogen gasin the presence of acetic acid.

In one embodiment, the tetracycline compound is tetracycline,doxycycline, methacycline, minocycline, or sancyline. In anotherembodiment, the tetracycline compound is a tetracycline compounddescribed in, for example, WO 03/079983, WO 02/12170, WO 02/04407, WO02.04406, WO 02/04405, WO 02/04404, WO 01/74761, WO 03/079984, WO03/075857, WO 03/057169, WO 02/072545, WO 02/072506, U.S. Ser. No.10/619,653, U.S. Ser. No. 09/895,857; U.S. Ser. No. 09/895,812; U.S.Pat. No. 5,326,759; U.S. Pat. No. 5,328,902; U.S. Pat. No. 5,495,031;U.S. Pat. No. 5,495,018; U.S. Pat. No. 5,495,030; U.S. Pat. No.5,495,032; U.S. Pat. No. 5,512,553; U.S. Pat. No. 5,675,030; U.S. Pat.No. 5,843,925; U.S. Pat. No. 5,886,175; U.S. Pat. No. 6,165,999; U.S.Pat. No. 3,239,499; WO 95/22529; U.S. Pat. No. 5,064,821; U.S. Pat. No.5,589,470; U.S. Pat. No. 5,811,412, or U.S. Application 20040002481.

Tetracycline compounds of the invention can be synthesized using methodsand reactive agents known in the art to react with amine groups. Forexample, acid chlorides are reactive agents which react to givetetracycline compounds of the inventions with various R⁴ and R^(4′)substituents (Scheme 11). Other reactive agents which can be used tosynthesize the compounds of the invention with various R⁴ and R^(4′)substituents are sulfonyl halides, sulfonyl anhydrides, and isocyanates.Further reactive agents which can be used to synthesize the compounds ofthe invention with various R⁴ and R^(4′)°substituents are aldehydesreacted under reducing conditions (Scheme 12)

The term “alkyl” includes saturated aliphatic groups, includingstraight-chain alkyl groups (e.g., methyl, ethyl, propyl, butyl, pentyl,hexyl, heptyl, octyl, nonyl, decyl, etc.), branched-chain alkyl groups(isopropyl, tert-butyl, isobutyl, etc.), cycloalkyl (alicyclic) groups(cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl), alkylsubstituted cycloalkyl groups, and cycloalkyl substituted alkyl groups.The term alkyl further includes alkyl groups, which can further includeoxygen, nitrogen, sulfur or phosphorous atoms replacing one or morecarbons of the hydrocarbon backbone. In certain embodiments, a straightchain or branched chain alkyl has 6 or fewer carbon atoms in itsbackbone (e.g., C₁-C₆ for straight chain, C₃-C₆ for branched chain), andmore preferably 4 or fewer. Likewise, preferred cycloalkyls have from3-8 carbon atoms in their ring structure, and more preferably have 5 or6 carbons in the ring structure. The term C₁-C₆ includes alkyl groupscontaining 1 to 6 carbon atoms.

Moreover, the term alkyl includes both “unsubstituted alkyls” and“substituted alkyls”, the latter of which refers to alkyl moietieshaving substituents replacing a hydrogen on one or more carbons of thehydrocarbon backbone. Such substituents can include, for example,alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, cyano, amino (including alkyl amino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety. Cycloalkyls can be further substituted, e.g.,with the substituents described above. An “alkylaryl” or an “arylalkyl”moiety is an alkyl substituted with an aryl (e.g.,phenylmethyl(benzyl)). The term “alkyl” also includes the side chains ofnatural and unnatural amino acids.

The term “aryl” includes groups, including 5- and 6-membered single-ringaromatic groups that may include from zero to four heteroatoms, forexample, benzene, phenyl, pyrrole, furan, thiophene, thiazole,isothiaozole, imidazole, triazole, tetrazole, pyrazole, oxazole,isooxazole, pyridine, pyrazine, pyridazine, and pyrimidine, and thelike. Furthermore, the term “aryl” includes multicyclic aryl groups,e.g., tricyclic, bicyclic, e.g., naphthalene, benzoxazole,benzodioxazole, benzothiazole, benzoimidazole, benzothiophene,methylenedioxyphenyl, quinoline, isoquinoline, napthridine, indole,benzofuran, purine, benzofuran, deazapurine, or indolizine. Those arylgroups having heteroatoms in the ring structure may also be referred toas “aryl heterocycles”, “heterocycles,” “heteroaryls” or“heteroaromatics”. The aromatic ring can be substituted at one or morering positions with such substituents as described above, as forexample, halogen, hydroxyl, alkoxy, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,alkylaminoacarbonyl, arylalkyl aminocarbonyl, alkenylaminocarbonyl,alkylcarbonyl, arylcarbonyl, arylalkylcarbonyl, alkenylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate,phosphonato, phosphinato, cyano, amino (including alkyl amino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety. Aryl groups can also be fused or bridged withalicyclic or heterocyclic rings which are not aromatic so as to form apolycycle (e.g., tetralin).

The term “alkenyl” includes unsaturated aliphatic groups analogous inlength and possible substitution to the alkyls described above, but thatcontain at least one double bond.

For example, the term “alkenyl” includes straight-chain alkenyl groups(e.g., ethylenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl,octenyl, nonenyl, decenyl, etc.), branched-chain alkenyl groups,cycloalkenyl (alicyclic) groups (cyclopropenyl, cyclopentenyl,cyclohexenyl, cycloheptenyl, cyclooctenyl), alkyl or alkenyl substitutedcycloalkenyl groups, and cycloalkyl or cycloalkenyl substituted alkenylgroups. The term alkenyl further includes alkenyl groups which includeoxygen, nitrogen, sulfur or phosphorous atoms replacing one or morecarbons of the hydrocarbon backbone. In certain embodiments, a straightchain or branched chain alkenyl group has 6 or fewer carbon atoms in itsbackbone (e.g., C₂-C₆ for straight chain, C₃-C₆ for branched chain).Likewise, cycloalkenyl groups may have from 3-8 carbon atoms in theirring structure, and more preferably have 5 or 6 carbons in the ringstructure. The term C₂-C₆ includes alkenyl groups containing 2 to 6carbon atoms.

Moreover, the term alkenyl includes both “unsubstituted alkenyls” and“substituted alkenyls”, the latter of which refers to alkenyl moietieshaving substituents replacing a hydrogen on one or more carbons of thehydrocarbon backbone. Such substituents can include, for example, alkylgroups, alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl,phosphate, phosphonato, phosphinato, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino),acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyland ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,alkylaryl, or an aromatic or heteroaromatic moiety.

The term “alkynyl” includes unsaturated aliphatic groups analogous inlength and possible substitution to the alkyls described above, butwhich contain at least one triple bond.

For example, the term “alkynyl” includes straight-chain alkynyl groups(e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl,nonynyl, decynyl, etc.), branched-chain alkynyl groups, and cycloalkylor cycloalkenyl substituted alkynyl groups. The term alkynyl furtherincludes alkynyl groups which include oxygen, nitrogen, sulfur orphosphorous atoms replacing one or more carbons of the hydrocarbonbackbone. In certain embodiments, a straight chain or branched chainalkynyl group has 6 or fewer carbon atoms in its backbone (e.g., C₂-C₆for straight chain, C₃-C₆ for branched chain). The term C₂-C₆ includesalkynyl groups containing 2 to 6 carbon atoms.

Moreover, the term alkynyl includes both “unsubstituted alkynyls” and“substituted alkynyls”, the latter of which refers to alkynyl moietieshaving substituents replacing a hydrogen on one or more carbons of thehydrocarbon backbone. Such substituents can include, for example, alkylgroups, alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl,phosphate, phosphonato, phosphinato, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino),acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyland ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,alkylaryl, or an aromatic or heteroaromatic moiety.

Unless the number of carbons is otherwise specified, “lower alkyl” asused herein means an alkyl group, as defined above, but having from oneto five carbon atoms in its backbone structure. “Lower alkenyl” and“lower alkynyl” have chain lengths of, for example, 2-5 carbon atoms.

The term “acyl” includes compounds and moieties which contain the acylradical (CH₃CO—) or a carbonyl group. It includes substituted acylmoieties. The term “substituted acyl” includes acyl groups where one ormore of the hydrogen atoms are replaced by for example, alkyl groups,alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, cyano, amino (including alkyl amino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety.

The term “acylamino” includes moieties wherein an acyl moiety is bondedto an amino group. For example, the term includes alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido groups.

The term “aroyl” includes compounds and moieties with an aryl orheteroaromatic moiety bound to a carbonyl group. Examples of aroylgroups include phenylcarboxy, naphthyl carboxy, etc.

The terms “alkoxyalkyl”, “alkylaminoalkyl” and “thioalkoxyalkyl” includealkyl groups, as described above, which further include oxygen, nitrogenor sulfur atoms replacing one or more carbons of the hydrocarbonbackbone, e.g., oxygen, nitrogen or sulfur atoms.

The term “alkoxy” includes substituted and unsubstituted alkyl, alkenyl,and alkynyl groups covalently linked to an oxygen atom. Examples ofalkoxy groups include methoxy, ethoxy, isopropyloxy, propoxy, butoxy,and pentoxy groups. Examples of substituted alkoxy groups includehalogenated alkoxy groups. The alkoxy groups can be substituted withgroups such as alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl,phosphate, phosphonato, phosphinato, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino),acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyland ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,alkylaryl, or an aromatic or heteroaromatic moieties. Examples ofhalogen substituted alkoxy groups include, but are not limited to,fluoromethoxy, difluoromethoxy, trifluoromethoxy, chloromethoxy,dichloromethoxy, trichloromethoxy, etc.

The term “amine” or “amino” includes compounds where a nitrogen atom iscovalently bonded to at least one carbon or heteroatom. The termincludes “alkyl amino” which comprises groups and compounds wherein thenitrogen is bound to at least one additional alkyl group. The term“dialkyl amino” includes groups wherein the nitrogen atom is bound to atleast two additional alkyl groups. The term “arylamino” and“diarylamino” include groups wherein the nitrogen is bound to at leastone or two aryl groups, respectively. The term “alkylarylamino,”“alkylaminoaryl” or “arylaminoalkyl” refers to an amino group which isbound to at least one alkyl group and at least one aryl group. The term“alkaminoalkyl” refers to an alkyl, alkenyl, or alkynyl group bound to anitrogen atom which is also bound to an alkyl group.

The term “amide,” “amido” or “aminocarbonyl” includes compounds ormoieties which contain a nitrogen atom which is bound to the carbon of acarbonyl or a thiocarbonyl group. The term includes “alkaminocarbonyl”or “alkylaminocarbonyl” groups which include alkyl, alkenyl, aryl oralkynyl groups bound to an amino group bound to a carbonyl group. Itincludes arylaminocarbonyl and arylcarbonylamino groups which includearyl or heteroaryl moieties bound to an amino group which is bound tothe carbon of a carbonyl or thiocarbonyl group. The terms“alkylaminocarbonyl,” “alkenylaminocarbonyl,” “alkynylaminocarbonyl,”“arylaminocarbonyl,” “alkylcarbonylamino,” “alkenylcarbonylamino,”“alkynylcarbonylamino,” and “arylcarbonylamino” are included in term“amide.” Amides also include urea groups (aminocarbonylamino) andcarbamates (oxycarbonylamino).

The term “carbonyl” or “carboxy” includes compounds and moieties whichcontain a carbon connected with a double bond to an oxygen atom. Thecarbonyl can be further substituted with any moiety which allows thecompounds of the invention to perform its intended function. Forexample, carbonyl moieties may be substituted with alkyls, alkenyls,alkynyls, aryls, alkoxy, aminos, etc. Examples of moieties which containa carbonyl include aldehydes, ketones, carboxylic acids, amides, esters,anhydrides, etc.

The term “thiocarbonyl” or “thiocarboxy” includes compounds and moietieswhich contain a carbon connected with a double bond to a sulfur atom.

The term “ether” includes compounds or moieties which contain an oxygenbonded to two different carbon atoms or heteroatoms. For example, theterm includes “alkoxyalkyl” which refers to an alkyl, alkenyl, oralkynyl group covalently bonded to an oxygen atom which is covalentlybonded to another alkyl group.

The term “ester” includes compounds and moieties which contain a carbonor a heteroatom bound to an oxygen atom which is bonded to the carbon ofa carbonyl group. The term “ester” includes alkoxycarboxy groups such asmethoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl,pentoxycarbonyl, etc. The alkyl, alkenyl, or alkynyl groups are asdefined above.

The term “thioether” includes compounds and moieties which contain asulfur atom bonded to two different carbon or hetero atoms. Examples ofthioethers include, but are not limited to alkthioalkyls,alkthioalkenyls, and alkthioalkynyls. The term “alkthioalkyls” includecompounds with an alkyl, alkenyl, or alkynyl group bonded to a sulfuratom which is bonded to an alkyl group. Similarly, the term“alkthioalkenyls” and alkthioalkynyls” refer to compounds or moietieswherein an alkyl, alkenyl, or alkynyl group is bonded to a sulfur atomwhich is covalently bonded to an alkynyl group.

The term “hydroxy” or “hydroxyl” includes groups with an —OH or —O⁻.

The term “halogen” includes fluorine, bromine, chlorine, iodine, etc.The term “perhalogenated” generally refers to a moiety wherein allhydrogens are replaced by halogen atoms.

The terms “polycyclyl” or “polycyclic radical” refer to two or morecyclic rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, arylsand/or heterocyclyls) in which two or more carbons are common to twoadjoining rings, e.g., the rings are “fused rings”. Rings that arejoined through non-adjacent atoms are termed “bridged” rings. Each ofthe rings of the polycycle can be substituted with such substituents asdescribed above, as for example, halogen, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, alkoxycarbonyl, alkylaminoacarbonyl,arylalkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl,arylcarbonyl, arylalkyl carbonyl, alkenylcarbonyl, aminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano,amido, amino (including alkyl amino, dialkylamino, arylamino,diarylamino, and alkylarylamino), acylamino (includingalkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino,imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkyl, alkylaryl, or anaromatic or heteroaromatic moiety.

The term “heteroatom” includes atoms of any element other than carbon orhydrogen. Preferred heteroatoms are nitrogen, oxygen, sulfur andphosphorus.

The term “prodrug moiety” includes moieties which can be metabolized invivo to a hydroxyl group and moieties which may advantageously remainesterified in vivo. Preferably, the prodrugs moieties are metabolized invivo by esterases or by other mechanisms to hydroxyl groups or otheradvantageous groups. Examples of prodrugs and their uses are well knownin the art (See, e.g., Berge et al. (1977) “Pharmaceutical Salts”, J.Pharm. Sci. 66:1-19). The prodrugs can be prepared in situ during thefinal isolation and purification of the compounds, or by separatelyreacting the purified compound in its free acid form or hydroxyl with asuitable esterifying agent. Hydroxyl groups can be converted into estersvia treatment with a carboxylic acid. Examples of prodrug moietiesinclude substituted and unsubstituted, branch or unbranched lower alkylester moieties, (e.g., propionoic acid esters), lower alkenyl esters,di-lower alkyl-amino lower-alkyl esters (e.g., dimethylaminoethylester), acylamino lower alkyl esters (e.g., acetyloxymethyl ester),acyloxy lower alkyl esters (e.g., pivaloyloxymethyl ester), aryl esters(phenyl ester), aryl-lower alkyl esters (e.g., benzyl ester),substituted (e.g., with methyl, halo, or methoxy substituents) aryl andaryl-lower alkyl esters, amides, lower-alkyl amides, di-lower alkylamides, and hydroxy amides. Preferred prodrug moieties are propionoicacid esters and acyl esters.

It will be noted that the structure of some of the tetracyclinecompounds of this invention includes asymmetric carbon atoms. It is tobe understood accordingly that the isomers arising from such asymmetry(e.g., all enantiomers and diastereomers) are included within the scopeof this invention, unless indicated otherwise. Such isomers can beobtained in substantially pure form by classical separation techniquesand by stereochemically controlled synthesis. Furthermore, thestructures and other compounds and moieties discussed in thisapplication also include all tautomers thereof.

Methods for Treating Tetracycline Responsive States

The invention also pertains to methods for treating a tetracyclineresponsive states in subjects, by administering to a subject aneffective amount of a tetracycline compound of the invention (e.g., acompound of Formula I or II, or otherwise described herein), such thatthe tetracycline responsive state is treated.

The term “treating” includes curing as well as ameliorating at least onesymptom of the state, disease or disorder, e.g., the tetracyclinecompound responsive state.

The language “tetracycline compound responsive state” or “tetracyclineresponsive state” includes states which can be treated, prevented, orotherwise ameliorated by the administration of a tetracycline compoundof the invention. Tetracycline compound responsive states includebacterial, viral, and fungal infections (including those which areresistant to other tetracycline compounds), cancer (e.g., prostate,breast, colon, lung melanoma and lymph cancers and other disorderscharacheterized by unwanted cellular proliferation, including, but notlimited to, those described in U.S. Pat. No. 6,100,248), arthritis,osteoporosis, diabetes, and other states for which tetracyclinecompounds have been found to be active (see, for example, U.S. Pat. Nos.5,789,395; 5,834,450; 6,277,061 and 5,532,227, each of which isexpressly incorporated herein by reference). Compounds of the inventioncan be used to prevent or control important mammalian and veterinarydiseases such as diarrhea, urinary tract infections, infections of skinand skin structure, ear, nose and throat infections, wound infection,mastitis and the like. In addition, methods for treating neoplasms usingtetracycline compounds of the invention are also included (van derBozert et al., Cancer Res., 48:6686-6690 (1988)). In a furtherembodiment, the tetracycline responsive state is not a bacterialinfection. In another embodiment, the tetracycline compounds of theinvention are essentially non-antibacterial. For example,non-antibacterial tetracycline compounds of the invention may have MICvalues greater than about 4 μg/ml (as measured by assays known in theart and/or the assay given in Example 9).

Tetracycline compound responsive states also include inflammatoryprocess associated states (IPAS). The term “inflammatory processassociated state” includes states in which inflammation or inflammatoryfactors (e.g., matrix metalloproteinases (MMPs), nitric oxide (NO), TNF,interleukins, plasma proteins, cellular defense systems, cytokines,lipid metabolites, proteases, toxic radicals, adhesion molecules, etc.)are involved or are present in an area in aberrant amounts, e.g., inamounts which may be advantageous to alter, e.g., to benefit thesubject. The inflammatory process is the response of living tissue todamage. The cause of inflammation may be due to physical damage,chemical substances, micro-organisms, tissue necrosis, cancer or otheragents. Acute inflammation is short-lasting, lasting only a few days. Ifit is longer lasting however, then it may be referred to as chronicinflammation.

IPAF's include inflammatory disorders. Inflammatory disorders aregenerally characterized by heat, redness, swelling, pain and loss offunction. Examples of causes of inflammatory disorders include, but arenot limited to, microbial infections (e.g., bacterial and fungalinfections), physical agents (e.g., burns, radiation, and trauma),chemical agents (e.g., toxins and caustic substances), tissue necrosisand various types of immunologic reactions.

Examples of inflammatory disorders include, but are not limited to,osteoarthritis, rheumatoid arthritis, acute and chronic infections(bacterial and fungal, including diphtheria and pertussis); acute andchronic bronchitis, sinusitis, and upper respiratory infections,including the common cold; acute and chronic gastroenteritis andcolitis; acute and chronic cystitis and urethritis; acute and chronicdermatitis; acute and chronic conjunctivitis; acute and chronicserositis (pericarditis, peritonitis, synovitis, pleuritis andtendinitis); uremic pericarditis; acute and chronic cholecystis; acuteand chronic vaginitis; acute and chronic uveitis; drug reactions; insectbites; burns (thermal, chemical, and electrical); and sunburn.

Tetracycline compound responsive states also include NO associatedstates. The term “NO associated state” includes states which involve orare associated with nitric oxide (NO) or inducible nitric oxide synthase(iNOS). NO associated state includes states which are characterized byaberrant amounts of NO and/or iNOS. Preferably, the NO associated statecan be treated by administering tetracycline compounds of the invention.The disorders, diseases and states described in U.S. Pat. Nos.6,231,894; 6,015,804; 5,919,774; and 5,789,395 are also included as NOassociated states. The entire contents of each of these patents arehereby incorporated herein by reference.

Other examples of NO associated states include, but are not limited to,malaria, senescence, diabetes, vascular stroke, neurodegenerativedisorders (Alzheimer's disease & Huntington's disease), cardiac disease(reperfusion-associated injury following infarction), juvenile diabetes,inflammatory disorders, osteoarthritis, rheumatoid arthritis, acute,recurrent and chronic infections (bacterial, viral and fungal); acuteand chronic bronchitis, sinusitis, and respiratory infections, includingthe common cold; acute and chronic gastroenteritis and colitis; acuteand chronic cystitis and urethritis; acute and chronic dermatitis; acuteand chronic conjunctivitis; acute and chronic serositis (pericarditis,peritonitis, synovitis, pleuritis and tendonitis); uremic pericarditis;acute and chronic cholecystis; cystic fibrosis, acute and chronicvaginitis; acute and chronic uveitis; drug reactions; insect bites;burns (thermal, chemical, and electrical); and sunburn.

The term “inflammatory process associated state” also includes, in oneembodiment, matrix metalloproteinase associated states (MMPAS). MMPASinclude states characterized by aberrant amounts of MMPs or MMPactivity. These are also include as tetracycline compound responsivestates which may be treated using compounds of the invention.

Examples of matrix metalloproteinase associated states (“MMPAS's”)include, but are not limited to, arteriosclerosis, corneal ulceration,emphysema, osteoarthritis, multiple sclerosis (Liedtke et al., Ann.Neurol. 1998, 44:35-46; Chandler et al., J. Neuroimmunol. 1997,72:155-71), osteosarcoma, osteomyelitis, bronchiectasis, chronicpulmonary obstructive disease, skin and eye diseases, periodontitis,osteoporosis, rheumatoid arthritis, ulcerative colitis, inflammatorydisorders, tumor growth and invasion (Stetler-Stevenson et al., Annu.Rev. Cell Biol. 1993, 9:541-73; Tryggvason et al., Biochim. Biophys.Acta 1987, 907:191-217; Li et al., Mol. Carcinog. 1998, 22:84-89)),metastasis, acute lung injury, stroke, ischemia, diabetes, aortic orvascular aneurysms, skin tissue wounds, dry eye, bone and cartilagedegradation (Greenwald et al., Bone 1998, 22:33-38; Ryan et al., Curr.Op. Rheumatol. 1996, 8; 238-247). Other MMPAS include those described inU.S. Pat. Nos. 5,459,135; 5,321,017; 5,308,839; 5,258,371; 4,935,412;4,704,383, 4,666,897, and RE 34,656, incorporated herein by reference intheir entirety.

In another embodiment, the tetracycline compound responsive state iscancer. Examples of cancers which the tetracycline compounds of theinvention may be useful to treat include all solid tumors, i.e.,carcinomas e.g., adenocarcinomas, and sarcomas. Adenocarcinomas arecarcinomas derived from glandular tissue or in which the tumor cellsform recognizable glandular structures. Sarcomas broadly include tumorswhose cells are embedded in a fibrillar or homogeneous substance likeembryonic connective tissue. Examples of carcinomas which may be treatedusing the methods of the invention include, but are not limited to,carcinomas of the prostate, breast, ovary, testis, lung, colon, andbreast. The methods of the invention are not limited to the treatment ofthese tumor types, but extend to any solid tumor derived from any organsystem. Examples of treatable cancers include, but are not limited to,colon cancer, bladder cancer, breast cancer, melanoma, ovariancarcinoma, prostatic carcinoma, lung cancer, and a variety of othercancers as well. The methods of the invention also cause the inhibitionof cancer growth in adenocarcinomas, such as, for example, those of theprostate, breast, kidney, ovary, testes, and colon.

In an embodiment, the tetracycline responsive state of the invention iscancer. The invention pertains to a method for treating a subjectsuffering or at risk of suffering from cancer, by administering aneffective amount of a substituted tetracycline compound, such thatinhibition cancer cell growth occurs, i.e., cellular proliferation,invasiveness, metastasis, or tumor incidence is decreased, slowed, orstopped. The inhibition may result from inhibition of an inflammatoryprocess, down-regulation of an inflammatory process, some othermechanism, or a combination of mechanisms. Alternatively, thetetracycline compounds may be useful for preventing cancer recurrence,for example, to treat residual cancer following surgical resection orradiation therapy. The tetracycline compounds useful according to theinvention are especially advantageous as they are substantiallynon-toxic compared to other cancer treatments. In a further embodiment,the compounds of the invention are administered in combination withstandard cancer therapy, such as, but not limited to, chemotherapy.

Examples of tetracycline responsive states also include neurologicaldisorders which include both neuropsychiatric and neurodegenerativedisorders, but are not limited to, such as Alzheimer's disease,dementias related to Alzheimer's disease (such as Pick's disease),Parkinson's and other Lewy diffuse body diseases, senile dementia,Huntington's disease, Gilles de la Tourette's syndrome, multiplesclerosis, amylotrophic lateral sclerosis (ALS), progressivesupranuclear palsy, epilepsy, and Creutzfeldt-Jakob disease; autonomicfunction disorders such as hypertension and sleep disorders, andneuropsychiatric disorders, such as depression, schizophrenia,schizoaffective disorder, Korsakoff's psychosis, mania, anxietydisorders, or phobic disorders; learning or memory disorders, e.g.,amnesia or age-related memory loss, attention deficit disorder,dysthymic disorder, major depressive disorder, mania,obsessive-compulsive disorder, psychoactive substance use disorders,anxiety, phobias, panic disorder, as well as bipolar affective disorder,e.g., severe bipolar affective (mood) disorder (BP-1), bipolar affectiveneurological disorders, e.g., migraine and obesity. Further neurologicaldisorders include, for example, those listed in the American PsychiatricAssociation's Diagnostic and Statistical manual of Mental Disorders(DSM), the most current version of which is incorporated herein byreference in its entirety.

Other examples of tetracycline compound responsive states are describedin WO 03/005971A2 and U.S. Patent Application Publication No.20040214800, each incorporated herein by reference.

The language “in combination with” another therapeutic agent ortreatment includes co-administration of the tetracycline compound,(e.g., inhibitor) and with the other therapeutic agent or treatment,administration of the tetracycline compound first, followed by the othertherapeutic agent or treatment and administration of the othertherapeutic agent or treatment first, followed by the tetracyclinecompound. The other therapeutic agent may be any agent which is known inthe art to treat, prevent, or reduce the symptoms of an IPAS.Furthermore, the other therapeutic agent may be any agent of benefit tothe patient when administered in combination with the administration ofan tetracycline compound. In one embodiment, the cancers treated bymethods of the invention include those described in U.S. Pat. Nos.6,100,248; 5,843,925; 5,837,696; or 5,668,122, incorporated herein byreference in their entirety.

In another embodiment, the tetracycline compound responsive state isdiabetes, e.g., juvenile diabetes, diabetes mellitus, diabetes type I,or diabetes type II. In a further embodiment, protein glycosylation isnot affected by the administration of the tetracycline compounds of theinvention. In another embodiment, the tetracycline compound of theinvention is administered in combination with standard diabetictherapies, such as, but not limited to insulin therapy. In a furtherembodiment, the IPAS includes disorders described in U.S. Pat. Nos.5,929,055; and 5,532,227, incorporated herein by reference in theirentirety.

In another embodiment, the tetracycline compound responsive state is abone mass disorder. Bone mass disorders include disorders where asubjects bones are disorders and states where the formation, repair orremodeling of bone is advantageous. For examples bone mass disordersinclude osteoporosis (e.g., a decrease in bone strength and density),bone fractures, bone formation associated with surgical procedures(e.g., facial reconstruction), osteogenesis imperfecta (brittle bonedisease), hypophosphatasia, Paget's disease, fibrous dysplasia,osteopetrosis, myeloma bone disease, and the depletion of calcium inbone, such as that which is related to primary hyperparathyroidism. Bonemass disorders include all states in which the formation, repair orremodeling of bone is advantageous to the subject as well as all otherdisorders associated with the bones or skeletal system of a subjectwhich can be treated with the tetracycline compounds of the invention.In a further embodiment, the bone mass disorders include those describedin U.S. Pat. Nos. 5,459,135; 5,231,017; 5,998,390; 5,770,588; RE 34,656;5,308,839; 4,925,833; 3,304,227; and 4,666,897, each of which is herebyincorporated herein by reference in its entirety.

In another embodiment, the tetracycline compound responsive state isacute lung injury. Acute lung injuries include adult respiratorydistress syndrome (ARDS), post-pump syndrome (PPS), and trauma. Traumaincludes any injury to living tissue caused by an extrinsic agent orevent. Examples of trauma include, but are not limited to, crushinjuries, contact with a hard surface, or cutting or other damage to thelungs.

The invention also pertains to a method for treating acute lung injuryby administering a substituted tetracycline compound of the invention.

The tetracycline responsive states of the invention also include chroniclung disorders. The invention pertains to methods for treating chroniclung disorders by administering a tetracycline compound, such as thosedescribed herein. The method includes administering to a subject aneffective amount of a substituted tetracycline compound such that thechronic lung disorder is treated. Examples of chronic lung disordersinclude, but are not limited, to asthma, cystic fibrosis, and emphesema.In a further embodiment, the tetracycline compounds of the inventionused to treat acute and/or chronic lung disorders such as thosedescribed in U.S. Pat. Nos. 5,977,091; 6,043,231; 5,523,297; and5,773,430, each of which is hereby incorporated herein by reference inits entirety.

In yet another embodiment, the tetracycline compound responsive state isischemia, stroke, or ischemic stroke. The invention also pertains to amethod for treating ischemia, stroke, or ischemic stroke byadministering an effective amount of a substituted tetracycline compoundof the invention. In a further embodiment, the tetracycline compounds ofthe invention are used to treat such disorders as described in U.S. Pat.No. 6,231,894; 5,773,430; 5,919,775 or 5,789,395, incorporated herein byreference.

In another embodiment, the tetracycline compound responsive state is askin wound. The invention also pertains, at least in part, to a methodfor improving the healing response of the epithelialized tissue (e.g.,skin, mucusae) to acute traumatic injury (e.g., cut, burn, scrape,etc.). The method may include using a tetracycline compound of theinvention (which may or may not have antibacterial activity) to improvethe capacity of the epithelialized tissue to heal acute wounds. Themethod may increase the rate of collagen accumulation of the healingtissue. The method may also decrease the proteolytic activity in theepthithelialized tissue by decreasing the collagenolytic and/orgellatinolytic activity of MMPs. In a further embodiment, thetetracycline compound of the invention is administered to the surface ofthe skin (e.g., topically). In a further embodiment, the tetracyclinecompound of the invention used to treat a skin wound, and other suchdisorders as described in, for example, U.S. Pat. Nos. 5,827,840;4,704,383; 4,935,412; 5,258,371; 5,308,839, 5,459,135; 5,532,227; and6,015,804; each of which is incorporated herein by reference in itsentirety.

In yet another embodiment, the tetracycline compound responsive state isan aortic or vascular aneurysm in vascular tissue of a subject (e.g., asubject having or at risk of having an aortic or vascular aneurysm,etc.). The tetracycline compound may by effective to reduce the size ofthe vascular aneurysm or it may be administered to the subject prior tothe onset of the vascular aneurysm such that the aneurysm is prevented.In one embodiment, the vascular tissue is an artery, e.g., the aorta,e.g., the abdominal aorta. In a further embodiment, the tetracyclinecompounds of the invention are used to treat disorders described in U.S.Pat. Nos. 6,043,225 and 5,834,449, incorporated herein by reference intheir entirety.

Bacterial infections may be caused by a wide variety of gram positiveand gram negative bacteria. The compounds of the invention are useful asantibiotics against organisms which are resistant to other tetracyclinecompounds. The antibiotic activity of the tetracycline compounds of theinvention may be determined using the method discussed in Example 9, orby using the in vitro standard broth dilution method described in Waitz,J. A., National Commission for Clinical Laboratory Standards, DocumentM7-A2, vol. 10, no. 8, pp. 13-20, 2^(nd) edition, Villanova, Pa. (1990).

The tetracycline compounds may also be used to treat infectionstraditionally treated with tetracycline compounds such as, for example,rickettsiae; a number of gram-positive and gram-negative bacteria; andthe agents responsible for lymphogranuloma venereum, inclusionconjunctivitis, psittacosis. The tetracycline compounds may be used totreat infections of, e.g., K. pneumoniae, Salmonella, E. hirae, A.baumanii, B. catarrhalis, H. influenzae, P. aeruginosa, E. faecium, E.coli, S. aureus or E. faecalis. In one embodiment, the tetracyclinecompound is used to treat a bacterial infection that is resistant toother tetracycline antibiotic compounds. The tetracycline compound ofthe invention may be administered with a pharmaceutically acceptablecarrier.

The language “effective amount” of the compound is that amount necessaryor sufficient to treat or prevent a tetracycline compound responsivestate. The effective amount can vary depending on such factors as thesize and weight of the subject, the type of illness, or the particulartetracycline compound. For example, the choice of the tetracyclinecompound can affect what constitutes an “effective amount”. One ofordinary skill in the art would be able to study the aforementionedfactors and make the determination regarding the effective amount of thetetracycline compound without undue experimentation.

The invention also pertains to methods of treatment againstmicroorganism infections and associated diseases. The methods includeadministration of an effective amount of one or more tetracyclinecompounds to a subject. The subject can be either a plant or,advantageously, an animal, e.g., a mammal, e.g., a human.

In the therapeutic methods of the invention, one or more tetracyclinecompounds of the invention may be administered alone to a subject, ormore typically a compound of the invention will be administered as partof a pharmaceutical composition in mixture with conventional excipient,i.e., pharmaceutically acceptable organic or inorganic carriersubstances suitable for parenteral, oral or other desired administrationand which do not deleteriously react with the active compounds and arenot deleterious to the recipient thereof.

Pharmaceutical Compositions of the Invention

The invention also pertains to pharmaceutical compositions comprising atherapeutically effective amount of a tetracycline compound (e.g., acompound of Formula I or II or any other compound described herein) and,optionally, a pharmaceutically acceptable carrier.

The language “pharmaceutically acceptable carrier” includes substancescapable of being coadministered with the tetracycline compound(s), andwhich allow both to perform their intended function, e.g., treat orprevent a tetracycline responsive state. Suitable pharmaceuticallyacceptable carriers include but are not limited to water, saltsolutions, alcohol, vegetable oils, polyethylene glycols, gelatin,lactose, amylose, magnesium stearate, talc, silicic acid, viscousparaffin, perfume oil, fatty acid monoglycerides and diglycerides,petroethral fatty acid esters, hydroxymethyl-cellulose,polyvinylpyrrolidone, etc. The pharmaceutical preparations can besterilized and if desired mixed with auxiliary agents, e.g., lubricants,preservatives, stabilizers, wetting agents, emulsifiers, salts forinfluencing osmotic pressure, buffers, colorings, flavorings and/oraromatic substances and the like which do not deleteriously react withthe active compounds of the invention.

The tetracycline compounds of the invention that are basic in nature arecapable of forming a wide variety of salts with various inorganic andorganic acids. The acids that may be used to prepare pharmaceuticallyacceptable acid addition salts of the tetracycline compounds of theinvention that are basic in nature are those that form non-toxic acidaddition salts, i.e., salts containing pharmaceutically acceptableanions, such as the hydrochloride, hydrobromide, hydroiodide, nitrate,sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate,lactate, salicylate, citrate, acid citrate, tartrate, pantothenate,bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate,gluconate, glucaronate, saccharate, formate, benzoate, glutamate,methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonateand palmoate [i.e., 1,1′-methylene-bis-(2-hydroxy-3-naphthoate)] salts.Although such salts must be pharmaceutically acceptable foradministration to a subject, e.g., a mammal, it is often desirable inpractice to initially isolate a tetracycline compound of the inventionfrom the reaction mixture as a pharmaceutically unacceptable salt andthen simply convert the latter back to the free base compound bytreatment with an alkaline reagent and subsequently convert the latterfree base to a pharmaceutically acceptable acid addition salt. The acidaddition salts of the base compounds of this invention are readilyprepared by treating the base compound with a substantially equivalentamount of the chosen mineral or organic acid in an aqueous solventmedium or in a suitable organic solvent, such as methanol or ethanol.Upon careful evaporation of the solvent, the desired solid salt isreadily obtained. The preparation of other tetracycline compounds of theinvention not specifically described in the foregoing experimentalsection can be accomplished using combinations of the reactionsdescribed above that will be apparent to those skilled in the art.

The preparation of other tetracycline compounds of the invention notspecifically described in the foregoing experimental section can beaccomplished using combinations of the reactions described above thatwill be apparent to those skilled in the art.

The tetracycline compounds of the invention that are acidic in natureare capable of forming a wide variety of base salts. The chemical basesthat may be used as reagents to prepare pharmaceutically acceptable basesalts of those tetracycline compounds of the invention that are acidicin nature are those that form non-toxic base salts with such compounds.Such non-toxic base salts include, but are not limited to those derivedfrom such pharmaceutically acceptable cations such as alkali metalcations (e.g., potassium and sodium) and alkaline earth metal cations(e.g., calcium and magnesium), ammonium or water-soluble amine additionsalts such as N-methylglucamine-(meglumine), and the loweralkanolammonium and other base salts of pharmaceutically acceptableorganic amines. The pharmaceutically acceptable base addition salts oftetracycline compounds of the invention that are acidic in nature may beformed with pharmaceutically acceptable cations by conventional methods.Thus, these salts may be readily prepared by treating the tetracyclinecompound of the invention with an aqueous solution of the desiredpharmaceutically acceptable cation and evaporating the resultingsolution to dryness, preferably under reduced pressure. Alternatively, alower alkyl alcohol solution of the tetracycline compound of theinvention may be mixed with an alkoxide of the desired metal and thesolution subsequently evaporated to dryness.

The preparation of other tetracycline compounds of the invention notspecifically described in the foregoing experimental section can beaccomplished using combinations of the reactions described above thatwill be apparent to those skilled in the art.

The tetracycline compounds of the invention and pharmaceuticallyacceptable salts thereof can be administered via either the oral,parenteral or topical routes. In general, these compounds are mostdesirably administered in effective dosages, depending upon the weightand condition of the subject being treated and the particular route ofadministration chosen. Variations may occur depending upon the speciesof the subject being treated and its individual response to saidmedicament, as well as on the type of pharmaceutical formulation chosenand the time period and interval at which such administration is carriedout.

The pharmaceutical compositions of the invention may be administeredalone or in combination with other known compositions for treatingtetracycline responsive states in a subject, e.g., a mammal. Preferredmammals include pets (e.g., cats, dogs, ferrets, etc.), farm animals(cows, sheep, pigs, horses, goats, etc.), lab animals (rats, mice,monkeys, etc.), and primates (chimpanzees, humans, gorillas). Thelanguage “in combination with” a known composition is intended toinclude simultaneous administration of the composition of the inventionand the known composition, administration of the composition of theinvention first, followed by the known composition and administration ofthe known composition first, followed by the composition of theinvention. Any of the therapeutically composition known in the art fortreating tetracycline responsive states can be used in the methods ofthe invention.

The tetracycline compounds of the invention may be administered alone orin combination with pharmaceutically acceptable carriers or diluents byany of the routes previously mentioned, and the administration may becarried out in single or multiple doses. For example, the noveltherapeutic agents of this invention can be administered advantageouslyin a wide variety of different dosage forms, i.e., they may be combinedwith various pharmaceutically acceptable inert carriers in the form oftablets, capsules, lozenges, troches, hard candies, powders, sprays(e.g., aerosols, etc.), creams, salves, suppositories, jellies, gels,pastes, lotions, ointments, aqueous suspensions, injectable solutions,elixirs, syrups, and the like. Such carriers include solid diluents orfillers, sterile aqueous media and various non-toxic organic solvents,etc. Moreover, oral pharmaceutical compositions can be suitablysweetened and/or flavored. In general, the therapeutically-effectivecompounds of this invention are present in such dosage forms atconcentration levels ranging from about 5.0% to about 70% by weight.

For oral administration, tablets containing various excipients such asmicrocrystalline cellulose, sodium citrate, calcium carbonate, dicalciumphosphate and glycine may be employed along with various disintegrantssuch as starch (and preferably corn, potato or tapioca starch), alginicacid and certain complex silicates, together with granulation binderslike polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally,lubricating agents such as magnesium stearate, sodium lauryl sulfate andtalc are often very useful for tabletting purposes. Solid compositionsof a similar type may also be employed as fillers in gelatin capsules;preferred materials in this connection also include lactose or milksugar as well as high molecular weight polyethylene glycols. Whenaqueous suspensions and/or elixirs are desired for oral administration,the active ingredient may be combined with various sweetening orflavoring agents, coloring matter or dyes, and, if so desired,emulsifying and/or suspending agents as well, together with suchdiluents as water, ethanol, propylene glycol, glycerin and various likecombinations thereof. The compositions of the invention may beformulated such that the tetracycline compositions are released over aperiod of time after administration.

For parenteral administration (including intraperitoneal, subcutaneous,intravenous, intradermal or intramuscular injection), solutions of atherapeutic compound of the present invention in either sesame or peanutoil or in aqueous propylene glycol may be employed. The aqueoussolutions should be suitably buffered (preferably pH greater than 8) ifnecessary and the liquid diluent first rendered isotonic. These aqueoussolutions are suitable for intravenous injection purposes. The oilysolutions are suitable for intraarticular, intramuscular andsubcutaneous injection purposes. The preparation of all these solutionsunder sterile conditions is readily accomplished by standardpharmaceutical techniques well known to those skilled in the art. Forparenteral application, examples of suitable preparations includesolutions, preferably oily or aqueous solutions as well as suspensions,emulsions, or implants, including suppositories. Therapeutic compoundsmay be formulated in sterile form in multiple or single dose formatssuch as being dispersed in a fluid carrier such as sterile physiologicalsaline or 5% saline dextrose solutions commonly used with injectables.

Additionally, it is also possible to administer the compounds of thepresent invention topically when treating inflammatory conditions of theskin. Examples of methods of topical administration include transdermal,buccal or sublingual application. For topical applications, therapeuticcompounds can be suitably admixed in a pharmacologically inert topicalcarrier such as a gel, an ointment, a lotion or a cream. Such topicalcarriers include water, glycerol, alcohol, propylene glycol, fattyalcohols, triglycerides, fatty acid esters, or mineral oils. Otherpossible topical carriers are liquid petrolatum, isopropylpalmitate,polyethylene glycol, ethanol 95%, polyoxyethylene monolauriate 5% inwater, sodium lauryl sulfate 5% in water, and the like. In addition,materials such as anti-oxidants, humectants, viscosity stabilizers andthe like also may be added if desired.

For enteral application, particularly suitable are tablets, dragees orcapsules having talc and/or carbohydrate carrier binder or the like, thecarrier preferably being lactose and/or corn starch and/or potatostarch. A syrup, elixir or the like can be used wherein a sweetenedvehicle is employed. Sustained release compositions can be formulatedincluding those wherein the active component is protected withdifferentially degradable coatings, e.g., by microencapsulation,multiple coatings, etc.

In addition to treatment of human subjects, the therapeutic methods ofthe invention also will have significant veterinary applications, e.g.for treatment of livestock such as cattle, sheep, goats, cows, swine andthe like; poultry such as chickens, ducks, geese, turkeys and the like;horses; and pets such as dogs and cats. Also, the compounds of theinvention may be used to treat non-animal subjects, such as plants.

It will be appreciated that the actual preferred amounts of activecompounds used in a given therapy will vary according to the specificcompound being utilized, the particular compositions formulated, themode of application, the particular site of administration, etc. Optimaladministration rates for a given protocol of administration can bereadily ascertained by those skilled in the art using conventionaldosage determination tests conducted with regard to the foregoingguidelines.

In general, compounds of the invention for treatment can be administeredto a subject in dosages used in prior tetracycline therapies. See, forexample, the Physicians' Desk Reference. For example, a suitableeffective dose of one or more compounds of the invention will be in therange of from 0.01 to 100 milligrams per kilogram of body weight ofrecipient per day, preferably in the range of from 0.1 to 50 milligramsper kilogram body weight of recipient per day, more preferably in therange of 1 to 20 milligrams per kilogram body weight of recipient perday. The desired dose is suitably administered once daily, or severalsub-doses, e.g. 2 to 5 sub-doses, are administered at appropriateintervals through the day, or other appropriate schedule.

It will also be understood that normal, conventionally known precautionswill be taken regarding the administration of tetracyclines generally toensure their efficacy under normal use circumstances. Especially whenemployed for therapeutic treatment of humans and animals in vivo, thepractitioner should take all sensible precautions to avoidconventionally known contradictions and toxic effects. Thus, theconventionally recognized adverse reactions of gastrointestinal distressand inflammations, the renal toxicity, hypersensitivity reactions,changes in blood, and impairment of absorption through aluminum,calcium, and magnesium ions should be duly considered in theconventional manner.

Furthermore, the invention also pertains to the use of a tetracyclinecompound of formula I, II or any other compound described herein, forthe preparation of a medicament. The medicament may include apharmaceutically acceptable carrier and the tetracycline compound is aneffective amount, e.g., an effective amount to treat a tetracyclineresponsive state.

EXEMPLIFICATION OF THE INVENTION

Compounds of the invention may be made as described below and/or byusing literature techniques known to those of ordinary skill of the art.

Example 1 Synthesis of 4-Oximinominocycline

To a solution of minocycline 2HCL (29.3 g, 60.6 mmol) in DMF (300 mL)was a 1:1 solution of hydroxylamine:water (7.98 mL, 120 mmol). Thesolution was heated to 80° C. for 2 hours while exposed to air. Aftercooling to room temperature, the solution was diluted with water (2 L).The water solution was filtered through a plug of DVB resin eluting witha 500 mL gradient of acetonitrile and water (5%-10%-20%-50%). At the 50%gradient, the product elutes as a yellow solution. The solution wasconcentrated under reduced pressure and further dried under vacuum toafford 12 g as a yellow/orange solid in 45% yield.

Example 2 Synthesis of 4-Oximinosancycline

To a solution of sancycline hydrate (8.29 g, 20.0 mmol) in DMF (100 mL)was added a 1:1 solution of hydroxylamine:water (2.70 mL, 40.0 mmol).The solution was heated to 80° C. for 2 hours while exposed to air.After cooling to room temperature, the solution was diluted with water(2 L). The water solution was filtered through a plug of DVB resineluting with a 500 mL gradient of acetonitrile and water(5%-10%-20%-50%). At the 50% gradient, the product elutes as a yellowsolution. The solution was concentrated under reduced pressure andfurther dried under vacuum to afford 3.7 g as a yellow/orange solid in46% yield.

Example 3 Synthesis of 4-Aminominocycline

To a solution of 4-oximinominocycline (11.1 g, 25.0 mmol) in methanol(250 mL) and acetic acid (7.22 mL, 125 mmol) was added 5% palladium oncarbon. The solution was flushed with hydrogen and placed under vacuumfor three successive cycles. After the final cycle, the flask was placedunder 50 psi of hydrogen for 16 hours. After flushing the flask withnitrogen and placing it under vacuum for three successive cycles, thesolution was filtered through a plug of celite, while rinsing withmethanol. The solution was concentrated under reduced pressure to afforda thick oil. The oil was poured into isopropanol (1 L) with vigorousstirring. The resulting suspension was collected on a sintered funnelwhile rinsing with cold isopropanol. The product was further dried underhigh vacuum overnight to afford 7.6 g as a light brown solid in 71%yield.

Example 4 Synthesis of 4-Aminosancycline

To a solution of 4-oximinosancycline, (11.1 g, 25.0 mmol) in methanol(250 mL) and acetic acid (7.22 mL, 125 mmol) was added 5% palladium oncarbon. The solution was flushed with hydrogen and placed under vacuumfor three successive cycles. After the final cycle, the flask was placedunder 50 psi of hydrogen for 16 hours. After flushing the flask withnitrogen and placing it under vacuum for three successive cycles, thesolution was filtered through a plug of celite, while rinsing withmethanol. The solution was concentrated under reduced pressure to afforda thick oil. The oil was poured into isopropanol (1 L) with vigorousstirring. The resulting suspension was collected on a sintered funnelwhile rinsing with cold isopropanol. The product was further dried underhigh vacuum overnight to afford 7.6 g as a light brown solid in 71%yield.

Example 5 Synthesis of 4-Acylminocycline

To a solution of 4-aminominocycline (502 mg, 1.00 mmol) in DMPU (2.5 mL)and acetonitrile (2.5 mL) at room temperature was added 1.5 eq.acylating reagent. After monitoring by HPLC, the reaction was judgedcomplete after 3 hours. The solution was diluted with water (20 mL) andwas run through a small plug of DVB resin eluting with a gradient (100mL) of acetonitrile and water (5%-10%-20%-50%). The fraction containingthe product was concentrated under reduced pressure and further driedunder vacuum.

Example 6 Synthesis of 4-Acylsancycline

To a solution of 4-aminosancycline (502 mg, 1.00 mmol) in DMPU (2.5 mL)and acetonitrile (2.5 mL) at room temperature was added 1.5 eq.acylating reagent. After monitoring by HPLC, the reaction was judgedcomplete after 3 hours. The solution was diluted with water (20 mL) andwas run through a small plug of DVB resin eluting with a gradient (100mL) of acetonitrile and water (5%-10%-20%-50%). The fraction containingthe product was concentrated under reduced pressure and further driedunder vacuum.

Example 7 Synthesis of 4-Dialkylminocycline

To a solution of 4-aminominocycline (502 mg, 1.00 mmol) and 5 eq.aldehyde in methanol (10 mL) and triethylamine (1 mL) was added 5%palladium on carbon (350 mg). The solution was flushed with hydrogen andplaced under vacuum for three successive cycles. After the final cycle,the flask was placed under 20 psi of hydrogen for 12 hours. Afterflushing the flask with nitrogen and placing it under vacuum for threesuccessive cycles, the solution was filtered through a plug of celite,while rinsing with methanol. The solution was concentrated under reducedpressure, diluted with 0.5M HCl (20 mL) and loaded onto a plug of DVBresin. The water solution was run throught the plug of DVB resin elutingwith a 100 mL gradient of acetonitrile and water (5%-10%-20%-50%). Thefraction containing the product was concentrated under reduced pressureand further dried under vacuum.

Example 8 Synthesis of 4-Dialkylsancycline

To a solution of 4-aminosancycline (502 mg, 1.00 mmol) and 5 eq.aldehyde in methanol (10 mL) and triethylamine (1 mL) was added 5%palladium on carbon (350 mg). The solution was flushed with hydrogen andplaced under vacuum for three successive cycles. After the final cycle,the flask was placed under 20 psi of hydrogen for 12 hours. Afterflushing the flask with nitrogen and placing it under vacuum for threesuccessive cycles, the solution was filtered through a plug of celite,while rinsing with methanol. The solution was concentrated under reducedpressure, diluted with 0.5M HCl (20 mL) and loaded onto a plug of DVBresin. The water solution was run throught the plug of DVB resin elutingwith a 100 mL gradient of acetonitrile and water (5%-10%-20%-50%). Thefraction containing the product was concentrated under reduced pressureand further dried under vacuum.

Example 9 In Vitro Minimum Inhibitory Concentration (MIC) Assay

The following assay is used to determine the efficacy of tetracyclinecompounds against common bacteria. 2 mg of each compound is dissolved in100 μl of DMSO. The solution is then added to cation-adjusted MuellerHinton broth (CAMHB), which results in a final compound concentration of200 μg per ml. The tetracycline compound solutions are diluted to 50 μLvolumes, with a test compound concentration of 0.098 μg/ml. Opticaldensity (OD) determinations are made from fresh log-phase broth culturesof the test strains. Dilutions are made to achieve a final cell densityof 1×10⁶ CFU/ml. At OD=1, cell densities for different genera should beapproximately:

E. coli 1 × 10⁹ CFU/ml S. aureus 5 × 10⁸ CFU/ml Enterococcus sp. 2.5 ×10⁹ CFU/ml  

50 μl of the cell suspensions are added to each well of microtiterplates. The final cell density should be approximately 5×10⁵ CFU/ml.These plates are incubated at 35° C. in an ambient air incubator forapproximately 18 hr. The plates are read with a microplate reader andare visually inspected when necessary. The MIC is defined as the lowestconcentration of the tetracycline compound that inhibits growth.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, numerous equivalents to thespecific procedures described herein. Such equivalents are considered tobe within the scope of the present invention and are covered by thefollowing claims. The contents of all references, patents, and patentapplications cited throughout this application are hereby incorporatedby reference. The appropriate components, processes, and methods ofthose patents, applications and other documents may be selected for thepresent invention and embodiments thereof.

1. A compound of formula (I):

wherein: R^(2′) and R^(2″) are each independently hydrogen, alkyl,alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,alkylamino, arylalkyl, aryl, heterocyclic, heteroaromatic or a prodrugmoiety; R¹⁰, R¹¹ and R¹² are each independently hydrogen, alkyl, aryl,benzyl, arylalkyl, or a pro-drug moiety; R^(3′) is hydroxyl, hydrogen,or a pro-drug moiety; R⁴ is O; R^(4a) is hydrogen, alkyl, alkenyl,alkynyl, or aryl; R⁵ and R^(5′) are each independently hydroxyl,hydrogen, thiol, alkanoyl, aroyl, alkaroyl, aryl, heteroaromatic, alkyl,alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,alkylamino, arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy; R⁶ andR^(6′) are each independently hydrogen, methylene, absent, hydroxyl,halogen, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylthio,alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl; R⁷ ishydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl, alkynyl,aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, arylalkyl, amino,arylalkenyl, arylalkynyl, acyl, aminoalkyl, heterocyclic, thionitroso,or —(CH₂)₀₋₃(NR^(7c))₀₋₁C(═W′)WR^(7a); R⁸ is hydrogen, hydroxyl,halogen, thiol, nitro, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylthio,alkylsulfinyl, alkylsulfonyl, alkylamino, amino, arylalkenyl,arylalkynyl, acyl, aminoalkyl, heterocyclic, thionitroso, or—(CH₂)₀₋₃(NR^(8c))₀₋₁C(=E′)ER^(8a); R⁹ is hydrogen, hydroxyl, halogen,thiol, nitro, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylthio,alkylsulfinyl, alkylsulfonyl, arylalkyl, amino, arylalkenyl,arylalkynyl, acyl, aminoalkyl, heterocyclic, thionitroso, or—(CH₂)₀₋₃(NR^(9c))₀₋₁C(═Z′)ZR^(9a); R^(7a), R^(7b), R^(7c), R^(7d),R^(7e), R^(7f), R^(8a), R^(8b), R^(8c), R^(8d), R^(8e), R^(8f), R^(9a),R^(9b), R^(9c), R^(9d), R^(9e), and R^(9f) are each independentlyhydrogen, acyl, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl, heterocyclic,heteroaromatic or a prodrug moiety; R¹³ is hydrogen, hydroxy, alkyl,alkenyl, alkynyl, alkoxy, alkylthio, aryl, alkylsulfinyl, alkylsulfonyl,alkylamino, or an arylalkyl; E is CR^(8d)R^(8e), S, NR^(8b) or O; E′ isO, NR^(8f), or S; W is CR^(7d)R^(7e), S, NR^(7b) or O; W′ is O, NR^(7f),or S; X is CHC(R¹³Y′Y), C═CR¹³Y, CR^(6′)R⁶, S, NR⁶, or O; Y′ and Y areeach independently hydrogen, halogen, hydroxyl, cyano, sulfhydryl,amino, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl,alkylsulfonyl, alkylamino, or an arylalkyl; Z is CR^(9d)R^(9e), S,NR^(9b) or O; Z′ is O, S, or NR^(9f), or a pharmaceutically acceptablesalt, ester or enantiomer thereof.
 2. The compound of claim 1, whereinR^(2′), R³, R¹⁰, R¹¹, and R¹² are each hydrogen or a prodrug moiety; Xis CR⁶R^(6′); and R^(2″), R⁵, R^(5′), R⁶, and R^(6′) are each hydrogen.3. The compound of claim 1, wherein R⁵ and R^(5′) are each hydrogen andX is CR⁶R^(6′), wherein R⁶ is methyl and R^(6′) is hydroxy.
 4. Thecompound of claim 1, wherein R⁵ is hydroxyl; X is CR⁶R^(6′); R⁶ ismethyl; and R^(5′) and R^(6′) are each hydrogen.
 5. The compound ofclaim 1, wherein X is CR⁶R^(6′); R⁵, R^(5′), R⁶ and R^(6′) are eachhydrogen and R⁷ is dimethylamino.
 6. The tetracycline compound of claim1, wherein said compound is:

or a pharmaceutically acceptable salt, ester or enantiomer thereof.
 7. Acompound of formula (II):

wherein: R^(2′) and R^(2″) are each independently hydrogen, alkyl,alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,alkylamino, arylalkyl, aryl, heterocyclic, heteroaromatic or a prodrugmoiety; R¹⁰, R¹¹ and R¹² are each independently hydrogen, alkyl, aryl,benzyl, arylalkyl, or a pro-drug moiety; R^(3′) is hydroxyl, hydrogen,or a pro-drug moiety; R⁴ is NR^(4c)R^(4d); R^(4c) and R^(4d) are eachindependently hydrogen, sulfonyl, arylcarbonyl, arylaminocarbonyl,aryloxycarbonyl, alkylcarbonyl, alkylaminocarbonyl, alkyloxycarbonyl,acyl, alkyl, alkenyl, alkynyl, or aryl; R⁵ and R^(5′) are eachindependently hydroxyl, hydrogen, thiol, alkanoyl, aroyl, alkaroyl,aryl, heteroaromatic, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, alkyl carbonyloxy,or aryl carbonyloxy; R⁶ and R^(6′) are each independently hydrogen,methylene, absent, hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl,aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or anarylalkyl; R⁷ is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl,alkenyl, alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,arylalkyl, amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl,heterocyclic, thionitroso, or —(CH₂)₀₋₃(NR^(7c))₀₋₁CC(═W′)WR^(7a); R⁸ ishydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl, alkynyl,aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino,amino, arylalkenyl, arylalkynyl, acyl, aminoalkyl, heterocyclic,thionitroso, or —(CH₂)₀₋₃(NR^(8c))₀₋₁C(=E′)ER^(8a); R⁹ is hydrogen,hydroxyl, halogen, thiol, nitro, alkyl, alkenyl, alkynyl, aryl, alkoxy,alkylthio, alkylsulfinyl, alkylsulfonyl, arylalkyl, amino, arylalkenyl,arylalkynyl, acyl, aminoalkyl, heterocyclic, thionitroso, or—(CH₂)₀₋₃(NR^(9c))₀₋₁C(═Z′)ZR^(9a); R^(7a), R^(7b), R^(7c), R^(7d),R^(7e), R^(7f), R^(8a), R^(8b), R^(8c), R^(8d), R^(8e), R^(8f), R^(9a),R^(9b), R^(9c), R^(9d), R^(9e), and R^(9f) are each independentlyhydrogen, acyl, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl, heterocyclic,heteroaromatic or a prodrug moiety; R¹³ is hydrogen, hydroxy, alkyl,alkenyl, alkynyl, alkoxy, alkylthio, aryl, alkylsulfinyl, alkylsulfonyl,alkylamino, or an arylalkyl; E is CR^(8d)R^(8e), S, NR^(8b) or O; E′ isO, NR^(8f), or S; W is CR^(7d)R^(7e), S, NR^(7b) or O; W′ is O, NR^(7f),or S; X is CHC(R¹³Y′Y), C═CR¹³Y, CR^(6′)R⁶, S, NR⁶, or O; Y′ and Y areeach independently hydrogen, halogen, hydroxyl, cyano, sulfhydryl,amino, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl,alkylsulfonyl, alkylamino, or an arylalkyl; Z is CR^(9d)R^(9e), S,NR^(9b) or O; Z′ is O, S, or NR^(9f), or a pharmaceutically acceptablesalt, ester or enantiomer thereof.
 8. The compound of claim 7, whereinR^(2′), R³, R¹⁰, R¹¹, and R¹² are each hydrogen or a prodrug moiety; Xis CR⁶R^(6′); and R^(2″), R⁵, R^(5′), R⁶, and R^(6′) are each hydrogen.9. The compound of claim 7, wherein R⁵ and R^(5′) are each hydrogen andX is CR⁶R^(6′), wherein R⁶ is methyl and R^(6′) is hydroxy.
 10. Thecompound of claim 7, wherein R⁵ is hydroxyl; X is CR⁶R^(6′); R⁶ ismethyl; and R^(5′) and R^(6′) are each hydrogen.
 11. The compound ofclaim 7, wherein X is CR⁶R^(6′); R⁵, R^(5′), R⁶ and R^(6′) are eachhydrogen and R⁷ is dimethylamino.
 12. The compound of claim 7, whereinR⁹ is hydrogen.
 13. The compound of claim 7, wherein R⁹ is substitutedor unsubstituted alkyl.
 14. The compound of claim 13, wherein R⁹ isaminoalkyl.
 15. The compound of claim 7, wherein said compound isselected from the group consisting of:

and pharmaceutically acceptable salts, esters and enantiomers thereof.16. A method for treating a tetracycline responsive state in a subject,comprising administering to said subject an effective amount of acompound of claim 1, such that said subject is treated.
 17. The methodof claim 16, wherein said tetracycline responsive state is a bacterialinfection, a viral infection, or a parasitic infection.
 18. The methodof claim 16, wherein said subject is a human.
 19. The method of claim16, wherein said compound is administered with a pharmaceuticallyacceptable carrier.
 20. A pharmaceutical composition comprising atherapeutically effective amount of a compound of claim 1 and apharmaceutically acceptable carrier.